Transformation of Competent Bacterial Cells: Electroporation

Abstract

Electroporation has become a valuable technique for the transfer of DNA into eukaryotic and prokaryotic cells. It is currently the most efficient means of transferring DNA, with efficiencies 20 times higher than obtained with maximally competent cells prepared with calcium chloride (CaCl2). Chemical transformation using CaCl2 is generally used for routine transformation when the DNA sample is not limiting. Electroporation is particularly useful in applications where high transformation efficiencies are specifically required. In essence, fast growing cells are centrifuged and washed in ice‐cold water. They are placed in a cuvette with the DNA and a high‐voltage pulse is applied. Cells are then plated out and selected for on agar.

Keywords: electroporation; electroporator; transformation; competent cells

References

Calvin NM and Hanawalt PC (1988) High efficiency transformation of bacterial cells by electroporation. Journal of Bacteriology 170: 2796–2801.

Dower WJ, Miller JF and Ragsdale CW (1988) High efficiency transformation of E. coli by high voltage electroporation. Nucleic Acids Research 16: 6127–6145.

Miller JF, Dower WJ and Tompkins LS (1988) High voltage electroporation of bacteria: genetic transformation of Campylobacter jejuni with plasmid DNA. Proceedings of the National Academy of Sciences of the USA 85: 856–860.

Shigekawa K and Dower WJ (1988) Electroporation of eukaryotes and prokaryotes: a general approach to the introduction of macromolecules into cells. Biotechniques 6: 742–751.

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How to Cite close
Anthony, R(Jul 2003) Transformation of Competent Bacterial Cells: Electroporation. In: eLS. John Wiley & Sons Ltd, Chichester. http://www.els.net [doi: 10.1038/npg.els.0003749]